Funds are requested to purchase a Queue LH Fermentor that permits controlled growth of microorganisms and a New Brunswick CEPA Z41 centrifuge for harvesting cells. The fermentor and centrifuge will be placed in a newly renovated fermentation facility. Other equipment to be in this facility and the renovation will be paid for by Washington University. These equipment items will facilitate research in the Curtiss lab to establish the genetic and biochemical bases for pathogenicity of various bacterial infectious disease agents in the genera Salmonella, Shigella, Streptococcus and Mycobacterium using a diversity of genetic (cloning, gene fusions, transposon mutagenesis, gene transfer, etc.), biochemical, immunological and microscopic techniques. Information learned from these basic studies will be used to develop novel means for vaccination, improved methods for early diagnosis and novel methods to screen for new drugs that might be effective in treating infections caused by bacterial agents. A specific goal of the vaccine development work is to enhance the secretory immune system so as to protect against infectious disease agents which colonize on or invade through a mucosal surface. Dr. Beachy's group will use the facility to recover Beta-galactosidase fusion proteins to serve as antigens specified by cloned cDNA whose mRNAs are regulated during the development of soybean embryos. Dr. Elgin's group will produce by recombinant techniques large quantities of non-histone chromosomal binding proteins that will be evaluated for ability to regulate developmental genes in Drosophila. Dr. Goodenough will grow Chlamydomonas to isolate the agglutinin responsible for sexual recognition and the ciliary protein dynein in order to analyze mating at the molecular level and the mechanism of ciliary motility, respectively. Dr. Silver's lab will use the fermentor to recover large quantities of proteins specified by the genes of the mercury-resistance operon to permit detailed structure-function studies including crystallographic analysis. Dr. Varner will use recombinant DNA techniques to permit production and recovery of large amounts of the proline-containing precursor of the hydroxyproline-rich glyco-protein of plant cell walls in order to study synthesis of the prolyl hydroxylase as a stress response in plants.